Organ care solution for ex-vivo machine perfusion of donor lungs

ABSTRACT

An ex-vivo lung solution for machine perfusion of donor lungs on OCS. The solution may be mixed with whole blood or packed red blood cells to form the OCS lung perfusion solution.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 35 U.S.C. §119(e), ofprovisional application U.S. Ser. No. 61/475,524, filed on Apr. 14,2011, entitled, “ORGAN CARE SOLUTION FOR EX-VIVO MACHINE PERFUSION OFDONOR LUNGS”, the entire subject matter of which is incorporated hereinby reference. This application also incorporates by reference, theentirety of U.S. application Ser. No. 12/099,715, filed on Apr. 8, 2008,entitled, “SYSTEMS AND METHODS FOR EX VIVO LUNG CARE”.

TECHNICAL FIELD

The disclosure generally relates a perfusion solution for ex-vivo organcare. More particularly, the disclosure relates to a solution formachine perfusion of donor lungs on an organ care system (“OCS”) atphysiologic or near-physiologic conditions.

BACKGROUND

Current organ preservation techniques typically involve hypothermicstorage of the organ in a chemical perfusion solution. In the case ofthe lung, it is typically flushed with a cold preservation solution suchas Perfadex™ and then immersed in that same cold solution until it istransplanted. These techniques utilize a variety of cold preservationsolutions, none of which sufficiently protect the lungs from tissuedamage resulting from ischemia. Such injuries are particularlyundesirable when an organ, such as a lung, is intended to betransplanted from a donor into a recipient.

Using conventional approaches, tissue injuries increase as a function ofthe length of time an organ is maintained ex-vivo. For example, in thecase of a lung, typically it may be preserved ex-vivo for only about 6to about 8 hours before it becomes unusable for transplantation. As aresult, the number of recipients who can be reached from a given donorsite is limited, thereby restricting the recipient pool for a harvestedlung. Compounding the effects of cold ischemia, current coldpreservation techniques preclude the ability to evaluate and assess anorgan ex-vivo. Because of this, less-than-optimal organs may betransplanted, resulting in post-transplant organ dysfunction or otherinjuries, or resuscitatable organs may be turned down.

Prolonged and reliable ex-vivo organ care would also provide benefitsoutside the context of organ transplantation. For example, a patient'sbody, as a whole, can typically tolerate much lower levels of chemo-,bio- and radiation therapy than many particular organs. An ex-vivo organcare system would permit an organ to be removed from the body andtreated in isolation, reducing the risk of damage to other parts of thebody. Thus, there is a need to develop techniques and perfusionsolutions that do not require hypothermic storage of the organ andextend the time during which an organ can be preserved in a healthystate ex-vivo. Such techniques would improve transplant outcomes andenlarge potential donor and recipient pools.

SUMMARY

The disclosure provides improved methods, solutions, and systems relatedto ex-vivo organ care. In general, in one aspect, the disclosurefeatures a lung OCS solution for machine perfusion of donor lungs on OCSat near physiologic conditions. In another aspect, the disclosureincludes a system and method for perfusing one or more lungs ex-vivo foran extended period of time in a functional and viable state maintenancemode at near physiologic conditions. In another aspect the disclosureincludes a method of producing a solution for ex-vivo perfusion of adonor lung at near physiologic conditions.

The present disclosure describes an OCS lung perfusion solution that canbe used for machine perfusion of donor lungs on OCS. The solution mayinclude energy-rich perfusion nutrients, as well as a supply oftherapeutics, vasodilators, endothelial stabilizers, and/orpreservatives for reducing edema and providing endothelial support tothe lungs. In a preferred embodiment, the solution comprises: dextran40; sodium chloride; potassium chloride; magnesium sulfate anhydrate;disodium phosphate anhydrate; monopotassium phosphate; glucosemonohydrate; milrinone; nitroglycerin; insulin; a multi-vitamin (M.V.I.Adult® or equivalent); sodium bicarbonate; methylprednisolone(SoluMedrol® or equivalent); cefazolin; Ciprofloxacin; voriconazole. Thesolution is mixed with whole blood or packed red blood cells to form theOCS lung perfusion solution. The solution provides the components formaintaining a functional (e.g., under respiration) and viable lungex-vivo at near physiologic conditions.

According to certain embodiments, solutions with particular solutes andconcentrations are selected and proportioned to provide for the organ tofunction at physiologic or near physiologic conditions. For example,such conditions include maintaining organ function at or near aphysiological temperature and/or preserving an organ in a state thatpermits normal cellular metabolism, such as protein synthesis andincreasing colloid pressure, minimize lung edema and cell swelling.

In another embodiment, a method of perfusing a lung is featured. Themethod includes: positioning the lung in an ex-vivo perfusion circuit;circulating an OCS lung solution specifically for machine perfusion ofdonor lungs on OCS through the lung, the fluid entering the lung througha pulmonary artery interface and leaving the lung through a left atrialinterface; ventilating the lung by flowing a ventilation gas through atracheal interface; deoxygenating the perfusion solution until apredetermined first value of oxygen content in the perfusion solution isreached; reoxygenating the perfusion solution by ventilating the lungwith an oxygenation gas until a predetermined second value of oxygencontent in the perfusion solution is reached; and determining acondition of the lung based on a time taken for the lung to cause theoxygen content level in the perfusion solution to change from the firstvalue of oxygen content to the second value of oxygen content.

In another embodiment, a method of producing a solution for perfusing alung at near physiologic conditions is featured. This method includescombining pre-weighed raw materials including nutrients, colloids,hormones, steroids, buffers and vasodilators with water for injection(“WFI”) and mixed with heating until fully dissolved, monitoring the pHlevel of the resulting solution, allowing the solution to cool,filtering the cooled solution, dispensing the solution into a primarycontainer and sterilizing the filled container.

In another aspect, a lung care system is featured. The lung systemincludes: a single use disposable module including an interface adaptedto couple the single use disposable module with the multiple use modulefor electro-mechanical interoperation with the multiple use module; alung chamber assembly optionally having a first interface for allowing aflow of a lung OCS perfusion solution into the lung, a second interfacefor allowing ventilation of the lung with a ventilation gas, and a thirdinterface for allowing a flow of the perfusion solution away from thelung, the lung chamber assembly including a dual drain system forcarrying the flow of the perfusion solution away from the lung, the dualdrain system comprising a measurement drain for directing a part of theperfusion solution flow to a sensor of a perfusion solution gas contentand a main drain for receiving a remaining part of perfusion solutionflow; and an OCS lung perfusion solution specifically for machineperfusion of donor lungs on OCS.

BRIEF DESCRIPTION OF THE DRAWINGS

The following figures depict illustrative embodiments in which likereference numerals refer to like elements. These depicted embodimentsmay not be drawn to scale and are to be understood as being illustrativeand not as limiting.

FIG. 1 is a schematic diagram of the lung perfusion circuit of thedescribed embodiment.

FIG. 2 is an illustration of the organ care system drawn from a45-degree angle from the front view, according to the describedembodiment.

FIG. 3 is an illustration of the lung perfusion module, according to thedescribed embodiment.

FIG. 4 is an illustration of the pulmonary artery cannula, according tothe described embodiment.

FIG. 5 is an illustration of the tracheal cannula, according to thedescribed embodiment.

FIG. 6 is an exploded illustration of the lung chamber, according to thedescribed embodiment.

FIG. 7 is a schematic diagram of the described embodiment of a portableorgan care system including shows the gas-related components of the lungperfusion module.

DETAILED DESCRIPTION

The following description and the drawings illustrate embodimentssufficiently to enable those skilled in the art to practice them. Otherembodiments may incorporate structural, logical, electrical, process,and other changes. Examples merely typify possible variations.Individual components and functions are optional unless explicitlyrequired, and the sequence of operations may vary. Portions and featuresof some embodiments may be included in or substituted for those ofothers. The scope of embodiments encompasses the full ambit of theclaims and all available equivalents of those claims.

Improved approaches to ex-vivo organ care are provided. Moreparticularly, various embodiments are directed to improved methods andsolutions relating to maintaining a lung at or near normal physiologicconditions in an ex-vivo environment. As used herein, “physiologicaltemperature” is referred to as temperatures between about 25 degrees C.and about 37 degrees C. A preferred embodiment comprises a lung OCSperfusion solution that may be administered in conjunction with an organcare system to maintain a lung in an equilibrium state by circulating aperfusion solution through the lung's vascular system, while causing thelung to rebreath a gas having an oxygen content sufficient to met thelung's metabolic needs.

The embodiments allow a lung to be maintained ex-vivo for extendedperiods of time, such as, for example, 3-24 or more hours. Such extendedex-vivo maintenance times expand the pool of potential recipients fordonor lungs, making geographic distance between donors and recipientsless important. Extended ex-vivo maintenance times also provide the timeneeded for better genetic and HLA matching between donor organs andorgan recipients, increasing the likelihood of a favorable outcome. Theability to maintain the organ in a near physiologic functioningcondition also allows a clinician to evaluate the organ's functionex-vivo, and identify organs that are damaged. This is especiallyvaluable in the case of the lung, since lungs are often compromised as adirect or indirect result of the cause of the death of the donor. Thuseven a newly harvested lung may be damaged. The ability to make a promptassessment of a harvested organ allows a surgeon to determine thequality of a lung and, if there is damage, to make a determination ofthe nature of the problem. The surgeon can then make a decision as towhether to discard the lung, or to apply therapy to the lung. Therapiescan include recruitment processes, removing or stapling off damagedareas of lung, suctioning secretions, cauterizing bleeding bloodvessels, and giving radiation treatment. The ability to assess and, ifnecessary provide therapy to lungs at several stages from harvesting toimplantation greatly improves the overall likelihood of lung transplantsuccess and increases the number of organs available for transplant. Insome instances, the improved assessment capability and extendedmaintenance time facilitates medical operators to perform physicalrepairs on donor organs with minor defects. Increased ex-vivo organmaintenance times can also provide for an organ to be removed from apatient, treated in isolation ex-vivo, and then put back into the bodyof a patient. Such treatment may include, without limitation,pharmaceutical treatments, gas therapies, surgical treatments, chemo-,bio-, gene and/or radiation therapies.

Overview of OCS Perfusion Solution

According to certain embodiments, a lung OCS perfusion solution withcertain solutes provides for the lungs to function at physiologic ornear physiologic conditions and temperature by supplying energy richnutrients, oxygen delivery, optimal oncotic pressure, pH and organmetabolism. The perfusion solution may also include therapeuticcomponents to help maintain the lungs and protect them against ischemia,reperfusion injury and other ill effects during perfusion. Therapeuticsmay also help mitigate edema, provide general endothelial tissue supportfor the lungs, and otherwise provide preventative or prophylactictreatment to the lungs.

The amounts of solutes provided describes preferred amounts relative toother components in the solution and may be scaled to providecompositions of sufficient quantity.

In one embodiment, the solution may include a phosphodiesteraseinhibitor. To improve gas exchange and diminish leukocytosis, anadenosine-3′,5′-cyclic monophosphate (cAMP) selective phosphodiesterasetype III (PDE III) inhibitor such as milrinone, aminone, anagrelide,bucladesine, cilostamide, cilostazol, enoximone, KMUP-1, quazinone,RPL-554, siguazodan, trequinsin, vesnarinone, zardaverine may be added.In a preferred embodiment milrinone is added. Milrinone has the effectsof vasorelaxation secondary to improved calcium uptake into thesarcoplasmic reticulum, inotropy (myocyte contraction) due tocAMP-mediated trans-sarcolemmal calcium flux, and lusitropy (myocyterelaxation) possibly due to improved actin-myosin complex dissociation.In a preferred embodiment milrinone is present in each 1 L of solutionin an amount of about 3400 mcg to about 4600. In a particularlypreferred embodiment, milrinone is present in each 1 L of solution in anamount of about 4000 mcg.

In certain embodiments the solution may include a nitrate which isuseful in the nitrogen cycle. Nitroglycerin is a nitrate that may beadded to the perfusion solution to promote stabilization of pulmonaryhemodynamics and improve arterial oxygenation after transplantation.When a lung is removed from the body, nitric oxide levels fall quicklybecause it is quenched by superoxide generated during reperfusion,resulting in damage to the lung tissue. Nitroglycerin can act to promotenitric oxide levels in a lung ex-vivo by way of intracellularS-nitrosothiol intermediates to directly stimulate guanylate cyclase orto release nitric oxide locally in effector cells. To this end,Nitroglycerin improves vascular homeostasis and improves organ functionby providing better arterial oxygenation after transplant. In apreferred embodiment nitroglycerin is present in each 1 L of solution inan amount of about 10 mg to about 50 mg.

In one other embodiment, magnesium sulfate anhydrate may be added to thesolution. Pulmonary artery blood pressure is lower than blood pressurein the rest of the body and in the case of pulmonary hypertension,magnesium sulfate promotes vasodilatation in constricted muscles of thepulmonary arteries by modulating calcium uptake, binding anddistribution in smooth muscle cells, thereby decreasing the frequency ofdepolarization of smooth muscle and thus promoting vasodilatation.Magnesium sulfate anhydrate is present in each 1 L of solution in anamount of about 0.083 g to about 0.1127 g. In a particularly preferredembodiment magnesium sulfate anhydrate is present in each 1 L ofsolution in an amount of about 0.098 g.

In a preferred embodiment, the addition of colloids offers numerousbenefits including improving erythrocyte deformability, preventingerythrocyte aggregation, inducing disbanding of already aggregated cellsand preserving endothelial-epithelial membrane. Colloids also haveanti-thrombotic effects by being able to coat endothelial surfaces andplatelets. In this embodiment dextran 40 is present in each 1 L ofsolution in an amount of about 42.5 g to about 57.5 g. In a particularlypreferred embodiment, dextran 40 is present in each 1 L of solution inan amount of about 50 g.

The solution may also contain electrolytes, such as sodium, potassium,chloride, sulfate, magnesium and other inorganic and organic chargedspecies, or combinations thereof. A suitable component may be thosewhere valence and stability permit, in an ionic form, in a protonated orunprotonated form, in salt or free base form, or as ionic or covalentsubstituents in combination with other components that hydrolyze andmake the component available in aqueous solutions. In this embodiment,sodium chloride is present in each 1 L of solution in an amount of about6.8 g to about 9.2 g. In a particularly preferred embodiment, sodiumchloride is present in each 1 L of solution in an amount of about 8 g.

In a preferred embodiment the solution may have a low-potassiumconcentration. A low-level of potassium results in improved lungfunction. A low potassium level may also protect the lung during highflow reperfusion and lead to a lower PA pressure and PVR, lower percentdecrease in dynamic airway compliance, and lower wet to dry ratio. Inthis embodiment potassium chloride is present in each 1 L of solution inan amount of about 0.34 g to about 0.46 g. In a particularly preferredembodiment potassium chloride is present in each 1 L of solution in anamount of about 0.4 g.

The solutions may include one or more energy-rich components to assistthe organ in conducting its normal physiologic function. Thesecomponents may include energy rich materials that are metabolizable,and/or components of such materials that an organ can use to synthesizeenergy sources during perfusion. Exemplary sources of energy-richmolecules include, for example, one or more carbohydrates. Examples ofcarbohydrates include glucose monohydrate, monosaccharides,disaccharides, oligosaccharides, polysaccharides, or combinationsthereof, or precursors or metabolites thereof. While not meant to belimiting, examples of monosaccharides suitable for the solutions includeoctoses; heptoses; hexoses, such as fructose, allose, altrose, glucose,mannose, gulose, idose, galactose, and talose; pentoses such as ribose,arabinose, xylose, and lyxose; tetroses such as erythrose and threose;and trioses such as glyceraldehyde. In a preferred embodiment glucosemonohydrate is present in each 1 L of solution an amount of about 1.7 gto about 2.3 g. In a particularly preferred embodiment glucosemonohydrate is present in each 1 L of solution an amount of about 2 g.

The solution may include other components to help maintain the organ andprotect it against ischemia, reperfusion injury and other ill effectsduring perfusion. In certain exemplary embodiments these components mayinclude a hormone to promote and regulate carbohydrate and fatmetabolism. Insulin acts to improve cell function by promoting optimumglucose and glycogen intake into the cells. In this preferred embodimenteach 1 L of the solution may contain about 17 IU insulin to about 23 IUinsulin. In a particularly preferred embodiment each 1 L of the solutionmay contain 20 IU insulin.

In addition, the solution may include a multi-vitamin that providesanti-oxidants and co-enzymes and helps maintain the body's normalresistance and repair processes. The multi-vitamin may include certainfat soluble vitamins such as Vitamins A, D, E, and K, and water solublevitamins such as Vitamin C, Niacinamide, Vitamins B₂, B₁, B₆, andDexpanthenol, as well as stabilizers and preservatives. In a preferredembodiment, each 1 L of the solution contains one unit vial of M.V.I.Adult® multi-vitamin. M.V.I. Adult® includes fat soluble vitamins suchas Vitamins A, D, E, and K, and water soluble vitamins such as VitaminC, Niacinamide, Vitamins B₂, B₁, B₆, and Dexpanthenol, as well asstabilizers and preservatives in an aqueous solution.

The solution may also include an anti-inflammatory agent such as aglucocorticoid steroid. Glucocorticoid steroids act as anti-inflamatoryagents by activating to the cell's glucocorticoid receptors which inturn up-regulate the expression of anti-inflammatory proteins in thenucleus and reduce the expression of pro-inflammatory proteins.Glucocorticoid steroids include methylprednisolone, hydrocortisone,cortisone acetate, prednisone, dexamethasone, betamethasone,triamcinolone, beclometasone, fludrocortisone acetate and aldosterone.In this preferred embodiment, each 1 L of the solution may contain about0.85 g mg to about 1.15 g methylprednisolone (SoluMedrol® orequivalent). In a particularly preferred embodiment, each 1 L of thesolution may contain 1 g methylprednisolone (SoluMedrol® or equivalent)

In addition the solution may contain buffers to maintain the solution atan optimal pH. These may include disodium phosphate anhydrate, aphysiologic balancing buffer or monopotassium phosphate to maintain theaverage pH of the solution during lung tissue perfusion. In thisembodiment disodium phosphate anhydrate is present in each 1 L ofsolution in an amount of about 0.039 g to about 0.052 g, and/ormonopotassium phosphate in an amount of about 0.053 g to about 0.072 g.In a particularly preferred embodiment, disodium phosphate anhydrate ispresent in an amount of 0.046 g, and/or monopotassium phosphate in anamount of 0.063 g. In some embodiments, the solution contains sodiumbicarbonate, potassium phosphate, or TRIS buffer. In a preferredembodiment the sodium bicarbonate is present in each 1 L of solution inan amount of about 12.75 mEq to about 17.25 mEq. In a particularlypreferred embodiment each 1 L of the solution may initially containabout 15 mEq sodium bicarbonate (5 mEq to each 500 mL bottle and 2-3bottles are used), and additional amounts may be added throughoutpreservation based on clinical judgment. For example, 20-40 mEq can beadded to the system as part of priming.

Other suitable buffers include 2-morpholinoethanesulfonic acidmonohydrate (MES), cacodylic acid, H₂CO₃/NaHCO₃ (pK_(a1)), citric acid(pK_(a3)), bis(2-hydroxyethyl)-imino-tris-(hydroxymethyl)-methane(Bis-Tris), N-carbamoylmethylimidino acetic acid (ADA),3-bis[tris(hydroxymethyl)methylamino]propane (Bis-Tris Propane)(pK_(a1)), piperazine-1,4-bis(2-ethanesulfonic acid) (PIPES),N-(2-Acetamido)-2-aminoethanesulfonic acid (ACES), imidazole,N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES),3-(N-morpholino)propanesulphonic acid (MOPS),NaH.sub.2PO.sub.4/Na.sub.2HPO.sub.4 (pK.sub.a2),N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid (TES),N-(2-hydroxyethyl)-piperazine-N′-2-ethanesulfonic acid (HEPES),N-(2-hydroxyethyl)piperazine-N′-(2-hydroxypropanesulfonic acid)(HEPPSO), triethanolamine, N-[tris(hydroxymethyl)methyl]glycine(Tricine), tris hydroxymethylaminoethane (Tris), glycineamide,N,N-bis(2-hydroxyethyl)glycine (Bicine), glycylglycine (pK_(a2)),N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), or acombination thereof.

The solution may contain an antimicrobial or antifungal agent to preventinfection. These may include bacteria and fungal antimicrobial agentsthat provide protection against both gram negative and gram positivebacteria. Suitable antimicrobial or antifungal agents include cefazolin,ciprofloxacin, and voriconazole or equivalent. In a preferredembodiment, cefazolin is present in each 1 L of solution in an amount ofabout 0.85 g to about 1.15 g, ciprofloxacin is present in each 1 L ofsolution in an amount of about 0.17 g to about 2.3 g, and voriconazoleis present in each 1 L of solution in an amount of about 0.17 g to about2.3 g. In a particularly preferred embodiment, cefazolin is present ineach 1 L of solution in an amount of about 1 g, ciprofloxacin is presentin each 1 L of solution in an amount of about 0.2 g, and voriconazole ispresent in each 1 L of solution in an amount of about 0.2 g.Alternatively the solution may contain any effective antimicrobial orantifungal agent.

The solutions are preferably provided at a physiological temperature andmaintained thereabout throughout perfusion and recirculation.

In a preferred embodiment the OCS lung perfusion solution comprises anutrient, a colloid, a vasodilator, a hormone and a steroid.

In another preferred embodiment the solution comprises a nutrientincluding Glucose monohydrate, sodium chloride, potassium chloride, amulti-vitamin including fat-soluble and water-soluble vitamins; acolloid including dextran 40; a hormone including insulin; a steroidincluding methylprednisolone; buffering agents including disodiumphosphate anhydrate, monopotassium phosphate and sodium bicarbonate;vasodilators including milrinone, nitroglycerin and magnesium sulfateanhydrate; antimicrobial or antifungal agents including cefazolin,ciprofloxacin, and voriconazole.

In another preferred embodiment the solution comprises an effectiveamount of dextran 40; sodium chloride; potassium chloride; magnesiumsulfate anhydrate; disodium phosphate anhydrate; monopotassiumphosphate; glucose monohydrate; milrinone; nitroglycerin; insulin; amulti-vitamin (M.V.I. Adult® or equivalent); sodium bicarbonate;methylprednisolone (SoluMedrol® or equivalent); cefazolin;ciprofloxacin; voriconazole.

In a preferred embodiment of the OCS lung perfusion solution, each 1 Lof solution includes, milrinone in an amount of about 4000 mcg;nitroglycerin in an amount of about 10-50 mg; dextran 40 in an amount ofabout 50 g; sodium chloride in an amount of about 8 g; potassiumchloride in an amount of about 0.4 g; magnesium sulfate anhydrate in anamount of about 0.098 g; disodium phosphate anhydrate in an amount ofabout 0.046 g; monopotassium phosphate in an amount of about 0.063 g;glucose monohydrate in an amount of about 2 g; insulin in an amount ofabout 20 IU; a multi-vitamin (M.V.I. Adult® or equivalent) in the amountof about 1 unit vial; sodium bicarbonate is initially present in anamount of about 15 mEq; methylprednisolone in an amount of about 1 g.

In a particularly preferred embodiment of the OCS lung perfusionsolution, each 1 L of solution includes, milrinone in an amount of about4000 mcg; nitroglycerin in an amount of about 10-50 mg; dextran 40 in anamount of about 50 g; sodium chloride in an amount of about 8 g;potassium chloride in an amount of about 0.4 g; magnesium sulfateanhydrate in an amount of about 0.098 g; disodium phosphate anhydrate inan amount of about 0.046 g; monopotassium phosphate in an amount ofabout 0.063 g; glucose monohydrate in an amount of about 2 g; insulin inan amount of about 20 IU; a multi-vitamin (M.V.I. Adult® or equivalent)in the amount of about 1 unit vial; sodium bicarbonate is initiallypresent in an amount of about 15 mEq; methylprednisolone in an amount ofabout 1 g; cefazolin in an amount of about 1 g; ciprofloxacin in anamount of about 0.2 g; voriconazole in an amount of about 0.2 g.

In certain embodiments, the perfusion solution is maintained andprovided to the lungs at a near physiologic temperature. According toone embodiment, the perfusion solution employs a blood product-basedperfusion solution to more accurately mimic normal physiologicconditions. The perfusion solution may be supplemented with cellularmedia. The cellular media may include a blood product, such as wholeblood, or packed red blood cells; allogenic packed red blood cells thatare leukocyte depleted/reduced; donor's whole blood that is leukocyteand platelet depleted/reduced; and/or human plasma to achievecirculating hematocrit of 15-30%.

Overview of Method of Producing a Solution for Perfusing a Lung at NearPhysiologic Temperature

In another aspect, a method of producing a solution for perfusing a lungat near physiologic temperature is provided. In a preferred method, thepre-weighed raw materials and WFI are added to a stainless steel mixingtank and mixed with heating until fully dissolved. The pH of theresulting solution is monitored and adjusted during the mixing processwith 1M hydrochloric acid (HCl). The solution is allowed to cool andthen filtered through a 0.2 μm filter and finally dispensed into aprimary container. The filled container is terminally sterilized withheat using a sterilization cycle that has been validated to achieve aSterility Assurance Level of 10⁻⁶. The raw materials in a preferredembodiment include a nutrient, a colloid, a vasodilator, a hormone and asteroid for perfusing a lung at near physiologic conditions.

In another preferred embodiment the raw materials include a nutrientincluding glucose monohydrate, sodium chloride, potassium chloride, amulti-vitamin including M.V.I. Adult® or equivalent; a colloid includingdextran 40; a hormone including insulin; a steroid includingmethylprednisolone; buffering agents including disodium phosphateanhydrate, monopotassium phosphate and sodium bicarbonate; vasodilatorsincluding milrinone, nitroglycerin and magnesium sulfate anhydrate; anantimicrobial or antifungal agent.

In another preferred embodiment the raw materials include dextran 40;sodium chloride; potassium chloride; magnesium sulfate anhydrate;disodium phosphate anhydrate; monopotassium phosphate; glucosemonohydrate; milrinone; nitroglycerin; insulin; a multi-vitamin (M.V.I.Adult® or equivalent); sodium bicarbonate; methylprednisolone(SoluMedrol® or equivalent); antimicrobial or antifungal agentsincluding cefazolin, ciprofloxacin, and voriconazole for perfusing alung at near physiologic conditions.

In a preferred embodiment, for each 1 L of solution, the raw materialsinclude milrinone in an amount of about 4000 mcg; nitroglycerin in anamount of about 10-50 mg; dextran 40 in an amount of about 50 g; sodiumchloride in an amount of about 8 g; potassium chloride in an amount ofabout 0.4 g; magnesium sulfate anhydrate in an amount of about 0.098 g;disodium phosphate anhydrate in an amount of about 0.046 g;monopotassium phosphate in an amount of about 0.063 g; glucosemonohydrate in an amount of about 2 g; insulin in an amount of about 20IU; a multi-vitamin (M.V.I. Adult® or equivalent) in the amount of about1 unit vial; sodium bicarbonate is initially present in an amount ofabout 15 mEq; methylprednisolone in an amount of about 1 g; anantimicrobial or antifungal agent.

In another particularly preferred embodiment, for each 1 L of solution,the raw materials include milrinone in an amount of about 4000 mcg;nitroglycerin in an amount of about 10-50 mg; dextran 40 in an amount ofabout 50 g; sodium chloride in an amount of about 8 g; potassiumchloride in an amount of about 0.4 g; magnesium sulfate anhydrate in anamount of about 0.098 g; disodium phosphate anhydrate in an amount ofabout 0.046 g; monopotassium phosphate in an amount of about 0.063 g;glucose monohydrate in an amount of about 2 g; insulin in an amount ofabout 20 IU; a multi-vitamin (M.V.I. Adult® or equivalent) in the amountof about 1 unit vial; sodium bicarbonate is initially present in anamount of about 15 mEq; methylprednisolone in an amount of about 1 g;cefazolin in an amount of about 1 g; ciprofloxacin in an amount of about0.2 g; voriconazole in an amount of about 0.2 g.

Overview of Method of Flushing an Organ with a Solution Between Excisefrom the Donor and Instrumentation on OCS

In another aspect, there is provided a method of flushing an organ witha solution between excise from the body and instrumentation on OCS. Inthis embodiment, to prepare a donor lung for surgical removal from thedonor's chest and to remove all old donor blood from the lung, the donorlung is flushed ante-grade using the pulmonary artery with the solutionuntil the temperature of the donor lung is in the range of about 0degrees C. to about 30 degrees C. Additionally, the solution may be usedfor retrograde flush of the lung using the pulmonary veins to remove anyblood clots remaining in the donor lung prior to surgical removal of thelung from the donor's chest, and to ensure adequate homogenousdistribution of flush solution to all lung segments. The lungs areventilated using a ventilator during both ante-grade and retro-gradeflushing to allow for homogenous distribution of the solution and toincrease the oxygen concentration in the donor lung alveoli to minimizethe impact of ischemia/reperfusion injury on the donor lung. Once theante-grade and retrograde flushing of the donor lung is completed, thelung will be removed surgically while inflated to minimize collapsing ofthe alveoli. Once the donor lung is fully removed from the donor body,it is ready to the next phase of OCS perfusion.

In one embodiment, the solution comprises an energy-rich perfusionnutrient, a colloid, a hormone, a buffer, magnesium sulfate anhydrate,and a nitrate. In another embodiment, the solution comprises dextran 40;sodium chloride; potassium chloride; magnesium sulfate anhydrate;disodium phosphate anhydrate; monopotassium phosphate; glucosemonohydrate; nitroglycerin.

In a particularly preferred embodiment each 1 L of solution forante-grade flush comprises dextran 40 in an amount of about 50 g; sodiumchloride in an amount of about 8 g; potassium chloride in an amount ofabout 0.4 g; magnesium sulfate anhydrate in an amount of about 0.098 g;disodium phosphate anhydrate in an amount of about 0.046 g;monopotassium phosphate in an amount of about 0.063 g; glucosemonohydrate in an amount of about 2 g; nitroglycerin in an amount ofabout 50 mg.

In another particularly preferred embodiment each 1 L of solution forretrograde flush comprises dextran 40 in an amount of about 50 g; sodiumchloride in an amount of about 8 g; potassium chloride in an amount ofabout 0.4 g; magnesium sulfate anhydrate in an amount of about 0.098 g;disodium phosphate anhydrate in an amount of about 0.046 g;monopotassium phosphate in an amount of about 0.063 g; glucosemonohydrate in an amount of about 2 g; nitroglycerin in an amount ofabout 10 mg.

Overview of Method of Machine Perfusion Using Lung OCS PerfusionSolution

In another aspect, a method for machine perfusion of a donor lung isprovided. The method includes perfusing the donor lung with a OCS lungperfusion solution comprising: dextran 40; sodium chloride; potassiumchloride; magnesium sulfate anhydrate; disodium phosphate anhydrate;monopotassium phosphate; glucose monohydrate; milrinone; nitroglycerin;insulin; at least two vitamins; sodium bicarbonate; methylprednisolone(SoluMedrol® or equivalent); a microbial or antifungal agent.

In a further aspect, the method includes perfusing the donor lung with aparticularly preferred OCS lung perfusion solution comprising for each 1L of solution: milrinone in an amount of about 4000 mcg; nitroglycerinin an amount of about 10-50 mg; dextran 40 in an amount of about 50 g;sodium chloride in an amount of about 8 g; potassium chloride in anamount of about 0.4 g; magnesium sulfate anhydrate in an amount of about0.098 g; disodium phosphate anhydrate in an amount of about 0.046 g;monopotassium phosphate in an amount of about 0.063 g; glucosemonohydrate in an amount of about 2 g; insulin in an amount of about 20IU; a multi-vitamin (M.V.I. Adult® or equivalent) in the amount of about1 unit vial; sodium bicarbonate is initially present in an amount ofabout 15 mEq; methylprednisolone in an amount of about 1 g; cefazolin inan amount of about 1 g; ciprofloxacin in an amount of about 0.2 g;voriconazole in an amount of about 0.2 g.

Overview of the Lung Perfusion Circuit

FIG. 1 illustrates an exemplary lung perfusion circuit which can be usedto circulate the perfusion solution noted above. The circuit is housedentirely within a lung perfusion module, and all its components may bedisposable. The organ care system (OCS) disclosure, U.S. applicationSer. No. 12/099,715, includes an exemplary embodiment of a lungperfusion circuit and is incorporated in its entirety by reference. LungOCS perfusion solution 250 is placed in a reservoir and then circulateswithin the perfusion circuit, passing through various components of lungperfusion module before passing through the vascular system of lungs404. Pump 226 causes perfusion solution 250 to flow around the lungperfusion circuit. It receives perfusion solution 250 from reservoir224, and pumps the solution through compliance chamber 228 to heater230. Compliance chamber 228 is a flexible portion of tubing that servesto refine the flow characteristics nature of pump 226. Heater 230replaces heat lost by perfusion solution 250 to the environment duringcirculation of the fluid. In the described embodiment, the heatermaintains perfusion solution 250 at or near the physiologic temperatureof 30-37 degrees C., and preferably at about 34 degrees C. After passingthrough heater 230, perfusion solution 250 flows into gas exchanger 402.Gas exchanger 402 allows gases to be exchanged between gas and perfusionsolution 250 via a gas-permeable, hollow fiber membrane. However, thegas exchanger has an effective gas exchange surface area of about 1square meter, which is only a fraction of the 50-100 square metereffective exchange area of the lungs. Thus gas exchanger 402 has only alimited gas exchange capability compared to the lungs. Blood gassolenoid valve 204 regulates the supply of gas into gas exchanger 402.Sampling/injection port 236 facilitates the removal of a sample or theinjection of a chemical just before perfusion solution 250 reaches thelungs. Perfusion solution then enters lungs 404 through cannulatedpulmonary artery 232. Flow probe 114 measures the rate of flow ofperfusion fluid 250 through the system. In the described embodiment,flow probe 114 is placed on the perfusate line as it leads towards thepulmonary artery. Pressure sensor 115 measures pulmonary arterialpressure at the point of entry of perfusion fluid 250 into the lungs. Inthe described embodiment, perfusion solution 250 is the lung OCSsolution described previously.

FIG. 2 is an overall view of OCS console 100 showing the single use,disposable lung perfusion module in a semi-installed position. Asbroadly indicated in FIG. 2, single use disposable lung perfusion moduleis sized and shaped to fit into OCS console 100, and to couple with it.Overall, the unit has a similar form to the organ care system describedin U.S. patent application Ser. No. 11/788,865. Removable lung perfusionmodule 400, is insertable into OCS console 100 by means of a pivotingmechanism that allows module 400 to slide into the organ console modulefrom the front, as shown in FIG. 2, and then pivot towards the rear ofthe unit. Clasp mechanism 2202 secures lung perfusion module 400 inplace. In alternative embodiments, other structures and interfaces oflung perfusion module 400 are used to couple the module with OCS 100.When secured in place, electrical and optical connections (not shown)provide power and communication between OCS console 100 and lungperfusion module 400. Details of the electrical and optical connectionsare described in U.S. patent application Ser. No. 11/246,013, filed onOct. 7, 2005, the specification of which is incorporated by referenceherein in its entirety. A key component of lung perfusion module 400 isorgan chamber 2204, which is described in detail below. Batterycompartments 2206 and maintenance gas cylinder 220 (not shown) arelocated in the base of the OCS console 100. OCS console 100 is protectedby removable panels, such as front panels 2208. Just below lungperfusion module are perfusion solution sampling ports 234 and 236.Mounted on top of OCS console 100 is OCS monitor 300.

FIG. 3 is a front view of lung perfusion module 400. Organ chamber 2204includes a removable lid 2820 and housing 2802. Sampling ports,including LA sampling port 234 and PA sampling port 236 are visiblebelow organ chamber 2802. Gas exchanger 402, bellows 418, and bellowsplate 2502 are also visible in the figure.

The circulation path of the perfusion solution, which was firstdescribed in connection with FIG. 2, in terms of the components of lungperfusion module 400 is now addressed. Mounted below organ chamber 2204are perfusion solution reservoir 224, which stores perfusion solution250. The perfusion solution exits through one-way inflow valve 2306,line 2702, and pump dome 2704 to pump 226 (not shown). The perfusionsolution is pumped through perfusion solution line 2404 throughcompliance chamber 228, and then to perfusion solution heater 230. Afterpassing through heater 230, the perfusion solution passes throughconnecting line 2706 to gas exchanger 402.

The pulmonary artery (PA) cannula connects the perfusion circuit withthe vascular system of lungs 404. An exemplary embodiment of a pulmonaryartery (PA) cannula is shown in FIG. 4. Referring to FIG. 4, single PAcannula 802 has single insertion tube 804 for insertion into a singlePA, and is used to cannulate the PA at a point before it branches to thetwo lungs. To connect the cannula to the pulmonary artery, insertiontube 804 is inserted into the PA, and the PA is secured onto the tubewith sutures. The tracheal cannula 700 is inserted into the trachea toprovide a means of connection between the lung perfusion module 400 gascircuit and the lungs. FIG. 5 illustrate an exemplary tracheal cannulae.Cannula 700 includes tracheal insertion portion 704 to which the tracheais secured with a cable tie, or by other means. The tracheal cannula maybe clamped at flexible portion 706 prior to instrumentation to seal offair flow in and out of the lungs 404. Also illustrated is an optionallocking nut 708.

The perfusion solution exits gas exchanger 402 through connecting line2708 to the interface with the pulmonary artery. After flowing throughthe lung and exiting via the pulmonary vein and the left atrium, theperfusion solution drains through from the base of organ chamber 2204,as described below. These drains feed the perfusion solution toreservoir 224, where the cycle begins again.

Having described OCS console 100 and lung perfusion module 400, we nowdescribe organ chamber 2204. FIG. 6 shows an exploded view of thecomponents of organ chamber 2204. Base 2802 of chamber 2204 is shapedand positioned within lung perfusion module 400 to facilitate thedrainage of the perfusion solution. Organ chamber 2204 has two drains,measurement drain 2804, and main drain 2806, which receives overflowfrom the measurement drain. Measurement drain 2804 drains perfusionsolution at a rate of about 0.5 l/min, considerably less than perfusionsolution 250 flow rate through lungs 404 of between 1.5 l/min and 4l/min. Measurement drain leads to oxygen probe 118, which measures SaO₂values, and then leads on to reservoir 224. Main drain 2806 leadsdirectly to reservoir 224 without oxygen measurement. Oxygen probe 118,which is a pulse oxymeter in the described embodiment, cannot obtain anaccurate measurement of perfusion solution oxygen levels unlessperfusion solution 250 is substantially free of air bubbles. In order toachieve a bubble-free column of perfusion solution, base 2802 is shapedto collect perfusion solution 250 draining from lungs 404 into a poolthat collects above drain 2804. The perfusion solution pool allows airbubbles to dissipate before the perfusion solution enters drain 2804.The formation of a pool above drain 2804 is promoted by wall 2808, whichpartially blocks the flow of perfusion solution from measurement drain2804 to main drain 2806 until the perfusion solution pool is largeenough to ensure the dissipation of bubbles from the flow. Main drain2806 is lower than measurement drain 2804, so once perfusion solutionoverflows the depression surrounding drain 2804, it flows around wall2808, to drain from main drain 2806. In an alternate embodiment of thedual drain system, other systems are used to collect perfusion solutioninto a pool that feeds the measurement drain. In some embodiments, theflow from the lungs is directed to a vessel, such as a small cup, whichfeeds the measurement drain. The cup fills with perfusion solution, andexcess blood overflows the cup and is directed to the main drain andthus to the reservoir pool. In this embodiment, the cup performs afunction similar to that of wall 2808 in the embodiment described aboveby forming a small pool of perfusion solution from which bubbles candissipate before the perfusion solution flows into the measurement drainon its way to the oxygen sensor.

Lungs 404 are supported by support surface 2810. The surface is designedto support lungs 404 without applying undue pressure, while anglinglungs 404 slightly downwards towards the lower lobes to promote easydrainage of the perfusion solution. Support surface includes drainagechannels 2812 to collect and channel perfusion solution issuing fromlungs 404, and to guide the perfusion solution towards drain 2814, whichfeeds perfusion solution directly to the blood pool for measurementdrain 2804. To provide additional support for the lungs, lungs 404 arewrapped with a polyurethane wrap (not shown) when placed on supportsurface 2810. The polyurethane wrap anchors lungs 404, helps keep thelungs in a physiologic configuration, and prevents the bronchi frombeing kinked and limiting the total volume of inflation. The wrapprovides a smooth surface for the exterior of the lung to interface withorgan chamber 2204, reducing the risk of the chamber applying excessivepressure on any part of lungs 404, which might cause undesirablehemorrhaging.

FIG. 7 is a schematic diagram of the described embodiment of a portableorgan care system including the gas-related components of the lungperfusion module. Controller 202 manages the release of maintenance andassessment gases by controlling the valves, gas selector switch 216, andventilator 214, thus implementing the preservation of the lungs inmaintenance mode, or the assessment of the lungs in one of theassessment modes. Blood gas solenoid valve 204 controls the amount ofgas flowing into blood gas exchanger 402. Airway pressure sensor 206samples pressure in the airway of lungs 404, as sensed through isolationmembrane 408. Relief valve actuator 207 is pneumatically controlled, andcontrols relief valve 412. The pneumatic control is carried out byinflating or deflating orifice restrictors that block or unblock the airpathway being controlled. This method of control allows completeisolation between the control systems in lung console module 200 and theventilation gas loop in lung perfusion module 400. Pneumatic control 208controls relief valve 207 and bellows valve actuator 210. Ventilator 214is a mechanical device with an actuator arm that causes bellows 418 tocontract and expand, which causes inhalation and exhalation of gas intoand out of lungs 404.

Use Models

An exemplary model for using the solution described above in the organcare system is described below.

The process of preparing the OCS perfusion module 400 forinstrumentation begins by producing the solution by the method ofproducing a solution for perfusing a lung at near physiologictemperature as described previously. About 800 ml to about 2000 ml ofthe OCS lung perfusion solution is then added into the Organ Care System(OCS) sterile perfusion module 400. The solution is then supplementedwith about 500 ml to about 1000 ml of cellular media. The cellular mediamay include one or combination of the following to achieve totalcirculating hematocrit concentration between 15-30%: typed allogenicpacked red blood cells (pRBCs) that is leukocytes depleted/reduce;donor's whole blood that is leukocyte and platelet depleted/reduced;and/or human plasma to achieve circulating hematocrit of 15-30%. The OCSdevice operates to circulate and mix the solution and cellular mediawhile warming and oxygenating the solution using a built in fluid warmerand gas exchanger 402. Once the solution is fully mixed, warmed andoxygenated, the pH of the solution will be adjusted using sodiumbicarbonate or other available buffer solution as needed. Once thesolution's hematocrit, temperature and pH levels reach an acceptablestate, the donor lung will be instrumented on OCS.

Once the solution is fully mixed, pH is adjusted to 7.35-7.45 andhematocrit is adjusted to 15-30%, the donor lung will be instrumented onOCS. To begin instrumentation, first set the flow rate of the OCS Pump226 to about 0.05 L/min. to ensure that perfusion solution does not exitthe PA line 233 prior to connecting the trachea cannula 700. Place thelung in the OCS' organ chamber 224 and connect the trachea cannula 700to the OCS trachea connector 710 and unclamp trachea cannula at section706. Then connect a PA pressure monitoring line with pressure sensor115, to the PA cannula 802. Trim the OCS' PA cannula 802 and prepare toconnect to the OCS PA line connector 231. Next, increase the OCS' pump226 flow to about 0.3 to about 0.4 L/min. so that a low-flow column ofsolution exits the PA line 233. Then remove any air from the lung byconnecting the lung PA cannula 802 to the OCS PA line connector 231 andgradually filling the PA cannula 802 with perfusion solution. Once anair-free column of solution is reached inside the PA cannula 802, sealthe connection between the PA cannula 802 and the OCS PA line connector231.

Next, gradually raise the OCS fluid warmer 230 temperature to 37 degreesC., and bring the perfusion solution temperature from about 32 degreesC. to about 37 degrees C. Then begin increasing the pump flow gradually,ensuring that pulmonary arterial pressure (“PAP”) remains below 20 mmHg,until pulmonary flow rate reaches a target flow rate of at least 1.5L/min. When the lung reaches a temperature of about 30 degrees C. toabout 32 degrees C., begin OCS ventilation by turning the OCS ventilator214 to “preservation” mode. The ventilator settings for instrumentationand preservation are specified in Table 1.

TABLE 1 Ventilator Settings (Instrumentation and Preservation) ParameterRequirement Tidal Volume (TV) = or <6 ml/kg Respiratory Rate (RR) 10breaths/min Positive End Expiratory 7-8 cm H₂O Pressure (PEEP) Note:decrease to 5 cm H₂O after confirming adequate inflation of lungs(within 2 hours) I:E Ratio 1:2-1:3 Peak Airway Pressure <25 cm H2O(PAWP)

Next, gradually increase the perfusion and ventilation rate for up toabout 30 minutes until reaching full ventilation and perfusion and allowventilation parameters to stabilize. Once ventilation parameters of thedonor lung on OCS have stabilized, wrap the lung to avoid over inflationinjury to the donor lung ex-vivo. The lung may also be wrapped during“pause preservation” before beginning ventilation. During preservationof lung on OCS, ventilation settings are maintained as described inTable 1, the mean PAP is maintained under about 20 mmHg, and the pumpflow is maintained at not less than about 1.5 L/min. Blood glucose,electrolytes and pH levels are monitored and adjusted within normalphysiologic ranges by additional injections. Lung oxygenation functionmay be assessed using the OCS lung system in addition to lungcompliance. In some instances it is desirable to provide therapy to thelung as described previously. Fiberoptic bronchoscopy may be performedfor the donor lung ex-vivo on the OCS device. Once preservation andassessment of the donor lung on the OCS system is complete, the lung iscooled and removed from the OCS system to be transplanted into therecipient.

Donor lung cooling may be achieved by first shutting off the OCSpulsatile pump 226 and flush the donor lung with about 3 liters ofperfusion solution at a temperature of about 0 degrees C. to about 15degrees C. while continuing ventilation on the OCS system. Once theflush is complete the trachea 700 and pulmonary artery 802 cannulae maybe disconnected from the OCS and the lung will be immersed in coldpreservation solution until it is surgically attached to the recipient(transplanted). Alternatively, the entire system circulating OCSsolution may be cooled down to 0 degrees C. to about 15 degrees C. usinga heat-exchanger and cooling device while the lung is being ventilatedon OCS. Once the target temperature of about 0 degrees C. to about 15 isachieved, the trachea 700 and pulmonary artery 802 cannulae will bedisconnected from the OCS and the lung will be immersed in coldpreservation solution until it is surgically attached to the recipient(transplanted).

The described system may utilize any embodiment of the lung OCSperfusion solution. In a preferred embodiment, the solution is mixedwith red blood cells and placed into a system reservoir for use in thesystem.

It is to be understood that while the invention has been described inconjunction with the various illustrative embodiments, the forgoingdescription is intended to illustrate and not limit the scope of theinvention, which is defined by the scope of the appended claims. Forexample, a variety of systems and/or methods may be implemented based onthe disclosure and still fall within the scope of the invention. Otheraspects, advantages, and modifications are within the scope of thefollowing claims. All references cited herein are incorporated byreference in their entirety and made part of this application.

1. An ex-vivo perfusion solution for machine perfusion of donor lungscomprising: an energy-rich perfusion nutrient, a colloid, a hormone, asteroid, a buffer, magnesium sulfate anhydrate, and at least one of aphosphodiesterase inhibitor and a nitrate.
 2. The solution of claim 1,wherein the phosphodiesterase inhibitor is a phosphodiesterase 3inhibitor.
 3. The solution of claim 1, wherein the phosphodiesteraseinhibitor is milrinone.
 4. The solution of claim 1, wherein the nitrateis nitroglycerin.
 5. The solution of claim 1, wherein the solutionincludes the phosphodiesterase inhibitor and the nitrate, and thephosphodiesterase inhibitor is milrinone and the nitrate isnitroglycerin.
 6. The solution of claim 5, wherein each liter ofsolution comprises milrinone in amount of about 4000 mcg andnitroglycerine in an amount of about 10 mg to 50 mg.
 7. The solution ofclaim 1, additionally comprising whole blood.
 8. The solution of claim1, additionally comprising red blood cells.
 9. The solution of claim 5,additionally comprising whole blood.
 10. The solution of claim 5,additionally comprising red blood cells.
 11. The solution of claim 5,wherein the nutrient includes glucose monohydrate, sodium chloride,potassium chloride, and M.V.I. Adult® multi-vitamin or equivalent; thecolloid includes dextran 40; the hormone includes insulin; the steroidincludes methylprednisolone; and the buffer includes disodium phosphateanhydrate, monopotassium phosphate and sodium bicarbonate.
 12. Thesolution of claim 6, wherein each liter of the solution comprisesdextran 40 in the amount of about 50 g; sodium chloride in an amount ofabout 8 g; potassium chloride in an amount of about 0.4 g; magnesiumsulfate anhydrate in an amount of about 0.098 g; disodium phosphateanhydrate in an amount of about 0.046 g; monopotassium phosphate in anamount of about 0.063 g; glucose monohydrate in an amount of about 2 g;insulin in an amount of about 20 IU; the multi-vitamin in an amount ofabout 1 unit vial; sodium bicarbonate in an amount of about 15 mEq;methylprednisolone in an amount of about 1 g.
 13. The solution of claim5, wherein the hormone comprises insulin.
 14. The solution of claim 5,wherein the hormone comprises about 20 IU insulin in each liter ofsolution.
 15. The solution of claim 5, wherein nutrient comprises amulti-vitamin and glucose monohydrate.
 16. The solution of claim 15wherein the multi-vitamin includes fat-soluble and water-solublevitamins.
 17. The solution of claim 5, wherein nutrient comprises about2 g glucose monohydrate in each liter of solution.
 18. The solution ofclaim 5 wherein the buffer comprises sodium bicarbonate.
 19. Thesolution of claim 5 wherein the buffer initially comprises about 15 mEqsodium bicarbonate in each liter of solution.
 20. The solution of claim5 wherein the steroid comprises a glucocorticoid steroid.
 21. Thesolution of claim 20 wherein the glucocorticoid steroid comprisesmethylprednisolone.
 22. The solution of claim 5 wherein the steroidcomprises 1 g methylprednisolone in each liter of solution.
 23. A methodof perfusing a donor lung at or near physiologic conditions comprising:flowing perfusion liquid through the lung, the perfusion liquid being ata physiologic temperature, the perfusion liquid comprising a nutrient, acolloid, a hormone, a steroid, a buffer, magnesium sulfate anhydrate, anantimicrobial agent and at least one of a phosphodiesterase inhibitorand a nitrate.
 24. The method of claim 23, wherein the nutrient includesglucose monohydrate, sodium chloride, potassium chloride, and amulti-vitamin; the colloid includes dextran 40; the hormone includesinsulin; the steroid includes methylprednisolone; the buffer includesdisodium phosphate anhydrate, monopotassium phosphate and sodiumbicarbonate; phosphodiesterase inhibitor includes milrinone, and thenitrate includes nitroglycerin.
 25. The method of claim 24, wherein eachliter of liquid comprises milrinone in an amount of about 4000 mcg;nitroglycerin in an amount of about 10 mg to 50 mg; dextran 40 in theamount of about 50 g; sodium chloride in an amount of about 8 g;potassium chloride in an amount of about 0.4 g; magnesium sulfateanhydrate in an amount of about 0.098 g; disodium phosphate anhydrate inan amount of about 0.046 g; monopotassium phosphate in an amount ofabout 0.063 g; glucose monohydrate in an amount of about 2 g; insulin inan amount of about 20 IU; the multi-vitamin in an amount of about 1 unitvial; sodium bicarbonate in an amount of about 15 mEq;methylprednisolone in an amount of about 1 g.
 26. The method of claim 25further comprising mixing the perfusion liquid with whole blood.
 27. Themethod of claim 25 further comprising mixing the perfusion liquid withred blood cells.
 28. The method of claim 25 further comprising mixingthe perfusion liquid with leukocyte-depleted whole blood.
 29. A methodof producing a solution for perfusing a lung at near physiologicconditions comprising the steps of: adding pre-weighed amounts ofdextran 40, sodium chloride, potassium chloride (KCL), magnesium sulfateanhydrate, disodium phosphate anydrate, monopotassium phosphate, glucosemonohydrate, milrinone, nitroglycerin, antimicrobial agents and water toa container to form a solution; mixing and heating the solution untilfully dissolved; monitoring the pH of the solution during mixing andadjusting the pH with 1M hydrochloric acid; allowing the solution tocool; filtering the solution; dispensing the solution into a primarycontainer; sterilizing the filled primary container with heat using asterilization cycle that has been validated to achieve a SterilityAssurance Level of 10⁻⁶.
 30. A method of producing a perfusion solutioncomprising combining pre-weighed amounts of a nutrient, a colloid, ahormone, a steroid, a buffer, magnesium sulfate anhydrate, and at leastone of a phosphodiesterase inhibitor and a nitrate to form a solutionfor perfusing a lung at near physiologic conditions.
 31. The method ofclaim 30 wherein the solution includes the phosphodiesterase inhibitorand the nitrate, and the nutrient includes glucose monohydrate, sodiumchloride, potassium chloride, and a multi-vitamin, wherein themulti-vitamin is selected from the group consisting of M.V.I. Adult® orequivalent; the colloid includes dextran 40; the hormone includesinsulin; the steroid includes methylprednisolone; buffer includesdisodium phosphate anhydrate, monopotassium phosphate and sodiumbicarbonate; the phosphodiesterase inhibitor includes milrinone, and thenitrate includes nitroglycerin.
 32. The method of claim 31, wherein eachliter of solution includes milrinone in an amount of about 4000 mcg;nitroglycerin in an amount of about 10 mg to 50 mg; dextran 40 in theamount of about 50 g; sodium chloride in an amount of about 8 g;potassium chloride in an amount of about 0.4 g; magnesium sulfateanhydrate in an amount of about 0.098 g; disodium phosphate anhydrate inan amount of about 0.046 g; monopotassium phosphate in an amount ofabout 0.063 g; glucose monohydrate in an amount of about 2 g; insulin inan amount of about 20 IU; the multi-vitamin in an amount of about 1 unitvial; sodium bicarbonate in an amount of about 15 mEq;methylprednisolone in an amount of about 1 g; the method furthercomprising mixing and heating the solution until fully dissolved;monitoring the pH of the solution during mixing and adjusting the pHwith 1M hydrochloric acid; allowing the solution to cool; filtering thesolution; dispensing the solution into a primary container; sterilizingthe filled primary container with heat using a sterilization cycle thathas been validated to achieve a Sterility Assurance Level of 10⁻⁶. 33.The method of claim 32 further comprising mixing the perfusion liquidwith whole blood.
 34. The method of claim 32 further comprising mixingthe perfusion liquid with red blood cells.
 35. The method of claim 32further comprising mixing the perfusion liquid with leukocyte-depletedwhole blood.
 36. A system for perfusing a donor lung in a lung perfusioncircuit at or near physiologic conditions comprising: a single usedisposable lung care module including an interface adapted forattachment to the single use module, and a lung chamber assembly havinga first interface for allowing a flow of a perfusion solution into thelung and a second interface for allowing ventilation of the lung with aventilation gas; and a drain system for draining a flow of perfusionsolution from the lung chamber assembly; and the perfusion solutionincluding dextran 40; sodium chloride; potassium chloride; magnesiumsulfate anhydrate; disodium phosphate anhydrate; monopotassiumphosphate; glucose monohydrate; milrinone; nitroglycerin; insulin; amulti-vitamin; sodium bicarbonate; and methylprednisolone.
 37. A methodof flushing a lung prior to preservation on an OCS comprising: flushinga donor lung prior to excising the lung from the donor's body with asolution comprising a nutrient, a colloid, a buffer, magnesium sulfateanhydrate, and a nitrate; excising the donor lung from the donor's body;placing the lung on an organ care system.
 38. The method of claim 37wherein the nutrient includes glucose monohydrate, sodium chloride andpotassium chloride; the colloid includes dextran 40; the buffer includesdisodium phosphate anhydrate and monopotassium phosphate; and thenitrate includes nitroglycerin.
 39. The method of claim 38, wherein eachliter of solution comprises nitroglycerin in an amount of about 10 mg to50 mg; dextran 40 in the amount of about 50 g; sodium chloride in anamount of about 8 g; potassium chloride in an amount of about 0.4 g;magnesium sulfate anhydrate in an amount of about 0.098 g; disodiumphosphate anhydrate in an amount of about 0.046 g; monopotassiumphosphate in an amount of about 0.063 g; glucose monohydrate in anamount of about 2 g.
 40. The solution of claim 1 further comprising anantimicrobial agent.
 41. The solution of claim 40, wherein theantimicrobial agent comprises at least one of cefazolin, ciprofloxacin,and voriconazole.
 42. The solution of claim 40 wherein each liter ofsolution comprises cefazolin in an amount of about 1 g, ciprofloxacin inan amount of about 0.2 g, and voriconazole in an amount of about 0.2 g.43. An ex-vivo perfusion solution for machine perfusion of donor lungscomprising: an energy-rich perfusion nutrient, a colloid, a hormone, asteroid, a buffer, magnesium sulfate anhydrate, at least one of aphosphodiesterase inhibitor and a nitrate, and an antimicrobial agent.44. The solution of claim 6, wherein each liter of the solution furthercomprises dextran 40 in the amount of about 50 g; sodium chloride in anamount of about 8 g; potassium chloride in an amount of about 0.4 g;magnesium sulfate anhydrate in an amount of about 0.098 g; disodiumphosphate anhydrate in an amount of about 0.046 g; monopotassiumphosphate in an amount of about 0.063 g; glucose monohydrate in anamount of about 2 g; insulin in an amount of about 20 IU; themulti-vitamin in an amount of about 1 unit vial; sodium bicarbonate inan amount of about 15 mEq; methylprednisolone in an amount of about 1 g;cefazolin in an amount of about 1 g; ciprofloxacin in an amount of about0.2 g; voriconazole in an amount of about 0.2 g.